大黄素对模拟冷缺血再灌注后肝细胞内钙离子浓度及细胞凋亡的影响

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1、大黄素对模拟冷缺血再灌注后肝细胞内钙离子浓度及细胞凋亡的影响作者：祁翔，吕毅，沈乃营，刘昌，刘学民，王博祁翔【摘要】目的研究中药大黄素对模拟冷缺血再灌注后肝细胞内钙离子浓度及细胞凋亡的影响。方法体外培养肝细胞株HL7702，随机分为对照组和大黄素处理组。对照组未予大黄素处理，大黄素处理组按100、10、1μmol/L分为高、中、低3个浓度组，建立模拟冷缺血再灌注模型，流式细胞技术检测各组细胞内钙离子浓度及细胞凋亡水平，分别检测各组细胞培养上清液乳酸脱氢酶水平。结果冷缺血8h再灌注6h后，高、中、低浓度大黄素处理组钙离子荧光强度分别为(24.12±0.51)、(26.35±1

2、.34)、(39.12±1.94)，均显著低于对照组的105.29±1.01(P<0.01)。高、中、低浓度大黄素处理组细胞凋亡率分别为(5.46±0.41)%、(10.64±0.64)%、(11.90±0.50)%，均显著低于对照组的(25.40±1.41)%(P<0.01)。高、中浓度大黄素处理组上清液LDH含量分别为(179.67±18.57)u/L、(198.83±14.22)u/L，显著低于对照组的(351.33±34.16)u/L(P<0.01)。结论大黄素可降低模拟冷缺血再灌注后的肝细胞内钙离子浓度，抑制细胞凋亡，减轻肝细胞损伤。【关键词】大黄

3、素；肝细胞；冷缺血再灌注；细胞内钙离子浓度；凋亡11　　ABSTRACT:ObjectiveToinvestigatetheeffectofEmodinonintracellularcalciumconcentration([Ca2+]i)andapoptosisofhepaticcellsaftersimulatedcoldischemiareperfusion.MethodsGlucoseoxygendeprivation,lowtemperature,subsequentreoxygenationandrewarmingwereusedtoinduceischemi

4、areperfusioninjurymodelinculturedhepaticcellswhichweredividedinto4groups:controlgroupandEmodintreatedgroup(100,10and1μmol/Lgroups).Theintracellularcalciumconcentration([Ca2+]i)andapoptosisrateweredeterminedbyflowcytometry(FCM)respectively;thecontentoflactatedehydrogenase(LDH)insupernatant

5、wastested.ResultsIntracellularcalciumfluorescenceintensityinEmodintreatedgroupsofhigh,mediumandlowdensitywas24.12±0.51,26.35±1.34and39.12±1.94,respectively,whichweresignificantlylowerthan105.29±1.01incontrolgroup(P<0.01).ApoptosisrateinEmodintreatedgroupsofhigh,mediumandlowdensitywas0.

6、0546±0.0041,0.1064±0.0064and0.1190±0.0050,respectively,whichweresignificantlylowerthan0.2540±0.0141incontrolgroup(P<0.01).LDHwas(198.83±14.22)u/Land(179.67±18.57)u/LinEmodintreatedgroupsofmediumandhighdensityrespectively,whichwere11significantlylowerthan(351.33±34.16)u/Lincontrolgroup(P

7、<0.01).ConclusionEmodincouldreduce[Ca2+]iandinhibitapoptosisofhepaticcellsaftersimulatedcoldischemiareperfusion,thusprotectinghepaticcellseffectively.　　KEYWORDS:Emodin;hepaticcell;ischemiareperfusioninjury;intracellularfreecalciumconcentration;apopt